Contaminations Occurring in Fungal PCR Assays-Reference-Cited by-同舟云学术

Contaminations Occurring in Fungal PCR Assays

Published:1999-04 Issue:4 Volume:37 Page:1200-1202
ISSN:0095-1137
Container-title:Journal of Clinical Microbiology
language:en
Short-container-title:J Clin Microbiol

Author:

Loeffler Juergen¹,Hebart Holger¹,Bialek Ralf²,Hagmeyer Lars¹,Schmidt Diethard¹,Serey Francois-Prâseth¹,Hartmann Matthias¹,Eucker Jan³,Einsele Hermann¹

Affiliation:

1. Universität Tuebingen, Medizinische Klinik und Poliklinik, Abteilung II,1 and

2. Institut für Tropenmedizin,272076 Tuebingen, and

3. Universitätsklinikum Charité, 10117 Berlin,3 Germany

Abstract

ABSTRACT Successful in vitro amplification of fungal DNA in clinical specimens has been reported recently. In a collaboration among five European centers, the frequency and risk of contamination due to airborne spore inoculation or carryover contamination in fungal PCR were analyzed. The identities of all contaminants were specified by cycle sequencing and GenBank analysis. Twelve of 150 PCR assays that together included over 2,800 samples were found to be contaminated (3.3% of the negative controls were contaminated during the DNA extraction, and 4.7% of the PCR mixtures were contaminated during the amplification process). Contaminants were specified as Aspergillus fumigatus , Saccharomyces cerevisiae , and Acremonium spp. Further analysis showed that commercially available products like zymolyase powder or 10× PCR buffer may contain fungal DNA. In conclusion, the risk of contamination is not higher in fungal PCR assays than in other diagnostic PCR-based assays if general precautions are taken.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

Link

https://journals.asm.org/doi/pdf/10.1128/JCM.37.4.1200-1202.1999

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